Disease evaluation support method and disease evaluation support device

Abstract

To support appropriate evaluation for a patient's disease.SOLUTION: A disease evaluation support method comprises: obtaining data of a drug efficacy evaluation test for a given drug using cells by three-dimensionally culturing some of diseased tissues of a target patient; obtaining data of multiple immunostaining of a tissue specimen prepared from the diseased tissue of the target patient; and displaying the result of the drug efficacy evaluation test and the analysis result of multiplex immunostaining side by side on a display. Consequently, it is possible to support appropriate evaluation of the target patient's disease.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] The present invention relates to a disease evaluation support method and a disease evaluation support device. [Background technology]

[0002] Even when various diseases are given the same diagnosis, their effectiveness in treatment often varies depending on the patient's own biological environment. Traditionally, in Japan, treatment guidelines have been established based on past evidence (the accumulation of a patient's clinical information and treatment results), and treatments are determined in accordance with these guidelines and clinical tests such as blood tests. However, due to technical and ethical constraints, the information that forms the basis of evidence is not sufficient to identify an individual's disease status and determine a treatment plan. For example, in the case of chemotherapy for cancer treatment, multiple options are available for many cancers, but the drug is often changed based on the therapeutic effect and adverse events after administration.

[0003] Because this situation places a physical burden on patients and increases treatment costs, efforts such as "patient stratification" and "personalized medicine," which accurately grasp an individual's biological environment and disease status and then provide optimal treatment based on that, have been gaining attention in recent years. One such technology is gene panel testing using next-generation sequencers in cancer treatment. This method involves dissolving resected tumor tissue and analyzing the expression levels of multiple genes. However, due to technical limitations and restrictions on applicable subjects, unfortunately, as of 2021, less than 10% of cases in Japan have led to treatment. Disadvantages of gene panel testing include the inability to directly examine drug effects, the inability to obtain cell-specific information, and the inability to obtain spatial information on cells.

[0004] As a method for directly examining the effects of drugs, a technique for examining drug effects by culturing cells three-dimensionally from patient-derived biomaterials is being studied (see, for example, Patent Document 1 and Non-Patent Document 1). By using this technique, it is expected that it will be possible to predict with high accuracy whether or not a drug is effective, although the patient's biological environment cannot be known.

[0005] In relation to other drawbacks of gene panel testing, such as "cell-by-cell information" and "spatial information," recent research has revealed that tumor heterogeneity and the tumor microenvironment are closely related to tumor characteristics and therapeutic efficacy, and multiple immunostaining methods have been developed as a method for analyzing these (see, for example, Patent Document 2 and Non-Patent Documents 2 to 4). Although multiple immunostaining methods cannot analyze a large number of genes like gene panel testing, they can obtain information at the single-cell level, including spatial and positional information, making it possible to obtain more detailed information about the state of diseased tissue (tumor tissue).

[0006] Patent Document 3 describes that the PD-L1 tumor status measured by immunohistochemistry (IHC) can be a predictive marker for identifying patients who are more likely to respond to cancer therapy using anti-PD-L1 antibody treatment. Non-Patent Documents 5 and 6 describe the observation of three-dimensional tumor spheroids in well plates using an imaging device. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Patent Publication No. 2021-164483 [Patent Document 2] International Publication No. 2017 / 087847 [Patent Document 3] Patent No. 6845840 [Non-patent literature]

[0008] [Non-Patent Document 1] Nicolas Boucherit et al., "3D Tumor Models and Their Use for the Testing of Immunotherapies", Frontiers in Immunology, December 2020, Volume 11, Article

Outdoor Tool2

Outdoor Tools3

Outdoor Tools 4

Direct Environment 5

[0009] Incidentally, appropriate evaluation of a patient's disease, including prediction of the efficacy of therapeutic drugs, is important for patient treatment. As mentioned above, when performing drug efficacy evaluation tests using three-dimensionally cultured cells, the culture process itself takes time, which limits the number of drugs that can be evaluated before treatment begins. Furthermore, since the state of the diseased tissue cannot be grasped, it is difficult to fully evaluate the patient's disease. On the other hand, while multiple immunostaining methods can grasp the state of diseased tissue, they cannot directly examine the effects of drugs, so they may be insufficient for evaluating the patient's disease. Therefore, in either case, the patient's disease may not be adequately evaluated.

[0010] The present invention has been made in view of the above problems, and aims to support appropriate evaluation of a patient's disease. [Means for solving the problem]

[0011] The invention of claim 1 is a disease evaluation support method, comprising: a) a step of obtaining data of a pharmacological efficacy evaluation test for a predetermined drug using cells obtained by three-dimensionally culturing a part of a diseased tissue of a subject patient; b) a step of obtaining data of multiple immunostaining of a tissue specimen prepared from the diseased tissue of the subject patient; and c) a step of displaying the results of the pharmacological efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display. The results of the drug efficacy evaluation test displayed on the display include a relationship between the concentration of the predetermined drug and the cell survival rate. .

[0012] The invention as set forth in claim 2 is the disease evaluation support method as set forth in claim 1, wherein the diseased tissue is cancer tissue.

[0013] The invention described in claim 3 is a disease evaluation support method described in claim 1 or 2, wherein the portion of the diseased tissue in step a) and the portion of the diseased tissue used to prepare the tissue specimen in step b) are adjacent to each other.

[0014] The invention of claim 4 is a disease evaluation support method, comprising: a) a step of obtaining data of a pharmacological efficacy evaluation test for a predetermined drug using three-dimensionally cultured blood circulating cancer cells of a subject patient; b) a step of obtaining data of multiple immunostaining of a tissue specimen prepared from cancer tissue, which is diseased tissue of the subject patient; and c) a step of displaying the results of the pharmacological efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display. The results of the drug efficacy evaluation test displayed on the display include a relationship between the concentration of the predetermined drug and the cell survival rate. .

[0015] The invention described in claim 5 is a disease evaluation support method described in any one of claims 1 to 4, further comprising a step of determining an evaluation value of the specified drug using the results of the drug efficacy evaluation test and the analysis results of the multiple immunohistochemistry, and displaying the evaluation value on the display.

[0016] The invention described in claim 6 is a disease evaluation support method described in any one of claims 1 to 5, wherein the results of the pharmacological evaluation test, the analysis results of the multiple immunostaining, treatment information, and treatment result information for a plurality of patients are prepared in advance, and the disease evaluation support method further comprises the step of identifying, from the plurality of patients, a patient whose results of the pharmacological evaluation test and the analysis results of the multiple immunostaining are similar to those of the target patient as a reference patient, and displaying the treatment information and the treatment result information of the reference patient on the display.

[0017] The invention described in claim 7 is a disease evaluation support method described in any one of claims 1 to 6, wherein in step a), the efficacy evaluation tests for multiple types of drugs are performed simultaneously using an imaging device.

[0018] The invention described in claim 8 is a disease evaluation support method described in any one of claims 1 to 7, wherein in step a), the efficacy evaluation test for multiple concentrations of the specified drug is performed simultaneously using an imaging device.

[0019] The invention of claim 9 is the disease evaluation support method of any one of claims 1 to 8, wherein the analysis results of the multiple immunostaining include analysis results of immune cells and / or tumor cells.

[0020] A tenth aspect of the present invention is the disease evaluation support method according to the ninth aspect, wherein the analysis results of the multiple immunostaining include the proportion or number of each type of immune cell.

[0021] The invention described in claim 11 is the disease evaluation support method described in claim 9 or 10, wherein the analysis results of the multiplex immunostaining include analysis results of immune cells in the tumor and various surrounding regions.

[0022] The invention of claim 12 is the disease evaluation support method of any one of claims 1 to 11, wherein the analysis results of the multiple immunostaining include a prediction of the effect of at least one type of drug.

[0023] The invention of claim 13 is a disease evaluation support device, comprising: a storage unit that stores data of a pharmacological efficacy evaluation test for a predetermined drug using cells obtained by three-dimensionally culturing a part of a diseased tissue of a subject patient, and data of multiple immunostaining of a tissue specimen prepared from the diseased tissue of the subject patient; and a display control unit that displays the results of the pharmacological efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display. The results of the drug efficacy evaluation test displayed on the display include a relationship between the concentration of the predetermined drug and the cell survival rate. .

[0024] The invention of claim 14 is a disease evaluation support device, comprising: a storage unit that stores data of a pharmacological efficacy evaluation test for a predetermined drug using three-dimensionally cultured blood circulating cancer cells of a subject patient, and data of multiple immunohistochemical staining of a tissue specimen prepared from cancer tissue, which is diseased tissue of the subject patient; and a display control unit that displays the results of the pharmacological efficacy evaluation test and the analysis results of the multiple immunohistochemical staining side by side on a display. The results of the drug efficacy evaluation test displayed on the display include a relationship between the concentration of the predetermined drug and the cell survival rate. . [Effects of the Invention]

[0025] According to the present invention, by displaying the results of a drug efficacy evaluation test and the results of multiplex immunostaining analysis side by side, it is possible to support appropriate evaluation of a patient's disease. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 1 is a diagram illustrating the configuration of a disease evaluation support device. [Figure 2] FIG. 1 illustrates the configuration of a computer. [Figure 3] FIG. 1 is a diagram showing a process flow for supporting disease evaluation of a subject patient. [Figure 4] FIG. 2 is a diagram illustrating an example of a display image on a display. [Figure 5] FIG. 10 shows another example of the analysis results of multiplex immunostaining. [Figure 6] FIG. 10 is a diagram showing an example of treatment result information. DETAILED DESCRIPTION OF THE INVENTION

[0027] FIG. 1 is a diagram showing the configuration of a disease evaluation support device 1 according to one embodiment of the present invention. In FIG. 1, components other than the disease evaluation support device 1 (a storage unit 40, an imaging device 41, and an immunostaining device 42) are also shown as blocks enclosed by dashed lines. The disease evaluation support device 1 is a device that supports doctors in evaluating patients' diseases. The disease evaluation support device 1 comprises a storage unit 21, a display control unit 22, a calculation unit 23, and a display 35. The disease evaluation support device 1 is realized by a computer executing calculation processing and the like in accordance with a predetermined program.

[0028] FIG. 2 is a diagram showing the configuration of the computer 3. The computer 3 has a typical computer system configuration including a CPU 31, a ROM 32, a RAM 33, a fixed disk 34, a display 35, an input unit 36, a reading device 37, a communication unit 38, a GPU 39, and a bus 30. The CPU 31 performs various arithmetic operations. The GPU 39 performs various arithmetic operations related to image processing. The ROM 32 stores basic programs. The RAM 33 stores various information. The fixed disk 34 stores information. The display 35 displays various information such as images. The input unit 36 ​​includes a keyboard 36a and a mouse 36b for receiving input from an operator. The reading device 37 reads information from a computer-readable recording medium 81 such as an optical disk, a magnetic disk, a magneto-optical disk, or a memory card. The communication unit 38 transmits and receives signals to and from external devices. The bus 30 is a signal circuit that connects the CPU 31, the GPU 39, the ROM 32, the RAM 33, the fixed disk 34, the display 35, the input unit 36, the reading device 37, and the communication unit 38.

[0029] In the computer 3, a program 811 is read in advance from the recording medium 81 via the reading device 37 and stored on the fixed disk 34. The program 811 may also be stored on the fixed disk 34 via a network. The computer 3 executes arithmetic processing and the like in accordance with the program 811, thereby realizing the disease evaluation support device 1 of FIG. 1. That is, the CPU 31, GPU 39, ROM 32, RAM 33, fixed disk 34 of the computer 3, and their peripheral components realize the disease evaluation support device 1. All or part of the functions of the disease evaluation support device 1 may be realized by dedicated electric circuits. Furthermore, these functions may be realized by multiple computers.

[0030] 1 stores data obtained by a drug efficacy evaluation test and multiple immunostaining (i.e., drug efficacy evaluation test data 211 and multiple immunostaining data 212), which will be described later. A calculation unit 23 performs analysis of the multiple immunostaining data, etc. A display control unit 22 displays the results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining on a display 35.

[0031] Next, a process for supporting the evaluation of a disease in a patient (hereinafter referred to as the "target patient") will be described with reference to FIG. 3. First, cells to be used for culture are prepared. In one example, diseased tissue of the target patient is partially removed by biopsy, surgery, or the like, and cells to be used for culture are collected from the diseased tissue. If the target patient's disease is cancer, circulating cancer cells from the target patient's blood may be collected for culture.

[0032] The collected cells are cultured three-dimensionally to obtain three-dimensional cultured cells. Three-dimensional cell culture is performed in a manner similar to the method described in JP 2021-164483 A (the above-mentioned Patent Document 1) or the method described in "3D Tumor Models and Their Use for the Testing of Immunotherapies" by Nicolas Boucherit et al. (Frontiers in Immunology, December 2020, Volume 11, Article 603640) (the above-mentioned Non-Patent Document 1). For example, a method of sequentially stacking cell layers is used. If the diseased tissue of the target patient is cancer tissue, the three-dimensional cultured cells contain cancer cells.

[0033] Next, a drug efficacy evaluation test for a predetermined drug is performed using the three-dimensional cultured cells. In this embodiment, multiple types of drugs are prepared, and multiple concentrations of each drug are changed and administered to the three-dimensional cultured cells in the wells of a well plate (e.g., a 96-well plate, a 386-well plate, etc.). That is, different types and concentrations of drugs are administered to the three-dimensional cultured cells in multiple wells. The administration of these drugs is carried out in a short period of time. The three-dimensional cultured cells may be cultured in the well plate, or may be placed in the well plate when the drugs are administered.

[0034] After a predetermined time has elapsed since the administration of the drug, the three-dimensional cultured cells in the multiple wells are observed using the imaging device 41. The imaging device 41 has an imaging unit and a movement mechanism. The movement mechanism sequentially positions the imaging unit above the multiple wells, thereby acquiring captured images of the three-dimensional cultured cells in each well. The imaging of the three-dimensional cultured cells in the multiple wells is completed in a short time. The calculation unit of the imaging device 41 calculates the fluorescence intensity of the three-dimensional cultured cells due to cell staining from the captured image, and determines the cell viability of the three-dimensional cultured cells based on the calculated fluorescence intensity. At this time, the three-dimensional cultured cells in each well are linked to the patient, drug type, drug concentration, etc., and the cell viability is also linked to the patient, drug type, drug concentration, etc. Data indicating the cell viability at multiple concentrations of multiple drugs is input into the disease evaluation support device 1 and stored as drug efficacy evaluation test data 211 in the memory unit 21 (step S11). In this manner, drug efficacy evaluation tests for multiple drugs at multiple concentrations are performed almost simultaneously using the imaging device 41. The cell viability of the three-dimensional cultured cells may be determined from bright-field photography and the spheroid size calculated from the bright-field image, as described in "High-throughput, bright-field imaging of 3D tumor spheroid growth and morphology for therapeutic and efficacy screening: A study using chemotherapeutic compounds" by Leena Mol Thuruthippallil et al. (American Association for Cancer Research, December 2015, Volume 14, Issue 12, Supplement 2, B75) (Non-Patent Document 5 mentioned above). Alternatively, the cell viability of the three-dimensional cultured cells may be determined by the calculation unit 23 of the disease evaluation support device 1. A drug efficacy evaluation test may be performed without using the imaging device 41.

[0035] Furthermore, a tissue specimen for multiple immunostaining is prepared from the diseased tissue of the subject patient. The diseased tissue is excised by biopsy, surgery, or the like. If the patient's disease is cancer, the diseased tissue is cancer tissue, and the tissue specimen includes cancer. If the cells used for the above-mentioned three-dimensional culture are collected from the diseased tissue, the tissue specimen for multiple immunostaining is also collected from the same diseased tissue. In this case, the site from which the cells are collected for three-dimensional culture and the site used to prepare the tissue specimen for multiple immunostaining can be collected from different regions of the diseased tissue, but it is preferable that the two sites are located close to each other. For example, if the diseased tissue of the subject patient is colon cancer, the distance between the two sites is preferably 10 cm or less. Alternatively, it is preferable that the two sites are included in tissue collected simultaneously with a biopsy needle or resected simultaneously with a scalpel. The tissue specimen can be prepared as a frozen section, but is preferably embedded in paraffin (to prepare a paraffin section) in order to preserve the morphological information of the cells and tissue.

[0036] Once the tissue specimen is prepared, multiplex immunostaining is performed. Multiplex immunostaining is performed, for example, in a manner similar to the method described in International Publication No. 2017 / 087847 (the above-mentioned Patent Document 2) or the method described in "Quantitative Multiplex Immunohistochemistry Reveals Myeloid-Inflamed Tumor-Immune Complexity Associated with Poor Prognosis" by Takahiro Tsujikawa et al. (Cell Reports, April 2017, Volume 19, pp. 203-217) (the above-mentioned Non-Patent Document 2). In this embodiment, the immunostaining device 42 repeatedly performs immunostaining and imaging on the tissue specimen while changing the type of antibody used. In one example, the tissue specimen is colored using a chromogenic substrate such as AEC or DAB, and images are acquired by bright-field observation using visible light. This allows multiple images corresponding to multiple types of proteins to be acquired. The data of the plurality of images is input from immunostaining apparatus 42 to disease evaluation support apparatus 1 and stored in storage unit 21 as multiplexed immunostaining data 212 (step S12).

[0037] Next, the display control unit 22 displays the results of the efficacy evaluation test and the analysis results of the multiple immunostaining side by side on the display 35 (step S13). FIG. 4 is a diagram showing an example of a display image 6 of the display 35. The display image 6 includes a first display area 61 and a second display area 62. The first display area 61 displays the results of the efficacy evaluation test, and the second display area 62 displays the analysis results of the multiple immunostaining. The results of the efficacy evaluation test are obtained from the efficacy evaluation test data 211. In the example of FIG. 4, a graph showing the relationship between the concentration of each drug and the cell viability and the concentration of the drug at which the cell viability is 50% (also called the 50% inhibitory concentration, hereinafter also referred to as "IC50") are displayed as the results of the efficacy evaluation test. The IC50 is a value indicating the efficacy of each drug, and the IC50 makes it possible to accurately predict the effect of the drug.

[0038] When displaying the analysis results of the multiplexed immunostaining, the calculation unit 23 analyzes the multiplexed immunostaining data 212. Various methods for analyzing the multiplexed immunostaining data 212 are known, including, for example, the method described in "Prognostic Significance of Spatial Immune Profiles in Human Solid Cancers" by Takahiro Tsujikawa et al. (Cancer Science, 2020, Volume 111, pp. 3426-3434) (Non-Patent Document 3) and the method described in "Spatial Profiles of Intratumoral PD-1" by Kanako Yoshimura et al. + Methods such as those described in "Helper T Cells Predict Prognosis in Head and Neck Squamous Cell Carcinoma" (Frontiers in Immunology, October 2021, Volume 12, Article 769534) (Non-Patent Document 4, above) can be used.

[0039] In the example of FIG. 4, an image showing the arrangement of tumor cells and multiple types of immune cells (e.g., T cells and B cells) and the proportions of the multiple types of immune cells in the tumor boundary region and within the tumor (excluding the boundary region) are obtained by analyzing the multiple immunostaining data 212 and displayed in the second display area 62. In the image at the top of the second display area 62 in FIG. 4, different colors are actually used for each type of cell. As described above, in the example of FIG. 4, the analysis results of the multiple immunostaining include the identification of immune cells and tumor cells, as well as the proportions of each type of immune cell in various regions of the tumor. This makes it possible to accurately grasp the cellular state of the patient's diseased tissue (here, tumor tissue). The second display area 62 in FIG. 4 may further display the proportions of each type of immune cell located around the tumor or outside the tumor, or the number of immune cells may be displayed instead of the proportions. The region of the image and the region from which the proportions of immune cells are calculated are set by the operator, etc.

[0040] The analysis results of multiple immunostaining may further distinguish whether or not each type of immune cell is in a state where its function is exerted (i.e., functional state) based on the presence or absence of expression of a specific protein. Furthermore, the analysis results of multiple immunostaining may not include identification of the cell type. Figure 5 shows, as another example of the second display area 62, a table showing the percentage of cells positive for multiple types of proteins (markers) and a two-dimensional histogram for each combination of two types of proteins.

[0041] Furthermore, the analysis results of multiple immunostaining may include a prediction of the effect of a drug. For example, the effect of an anti-PD-L1 antibody drug or the like can be predicted based on the positive rate in tumor cells by PD-L1 immunostaining (see, for example, Japanese Patent No. 6845840 (the above-mentioned Patent Document 3)). The drug whose effect is predicted by the analysis of multiple immunostaining may be the same as or different from the drug whose efficacy is being evaluated. There may be two or more types of drugs whose effect is predicted. It is preferable that the analysis results of multiple immunostaining include a prediction of the effect of at least one type of drug.

[0042] Next, the calculation unit 23 calculates an evaluation value for each type of drug that underwent the efficacy evaluation test using the results of the efficacy evaluation test and the analysis results of the multiplex immunostaining. The evaluation value is a value that predicts the efficacy of the drug in the target patient. In the following example of calculating the evaluation value, it is assumed that a population of one type of cell (hereinafter referred to as "target cells") is composed of cells positive for multiple predetermined proteins. The proteins of interest are denoted as "CDa," "CDb," and "CDc." To calculate the evaluation value, the proportion of CDa-positive cells (HTa), the proportion of CDb-positive cells (HTb), and the proportion of CDc-positive cells (HTc) are calculated among the target cells. Then, the evaluation value of drug A is calculated using the IC50 (50% inhibitory concentration) value DA of drug A, the IC50 value DB of drug B, and the IC50 value DC of drug C, according to Equation 1. α0 to α6 in Equation 1 are predetermined by collecting past medication result data and performing multivariate analysis.

[0043] (Number 1) Evaluation value of drug A = α0 + α1*HTa + α2*HTb + α3*HTc + α4*DA + α5*DB + α6*DC The evaluation values ​​for drugs B and C are calculated in the same manner, and the evaluation values ​​for drugs A, B, and C are displayed on the display 35 (step S14). The display of the evaluation values ​​does not necessarily have to be the display of the evaluation values ​​themselves. For example, of multiple categories preset for the numerical range in which the evaluation values ​​can be, a category containing the evaluation value of each drug may be identified, and the evaluation value of the drug may be substantially displayed by displaying a color or the like assigned to that category. The formula for calculating the evaluation value of a drug may be other than Equation 1. The display of the evaluation values ​​in step S14 may be performed by an operator's input instructing the display of the evaluation values ​​(the same applies to step S15 described below).

[0044] Furthermore, in disease evaluation support device 1, a patient whose pharmacological efficacy evaluation test results and multiple immunostaining analysis results are similar to those of the target patient is identified as a reference patient, and the treatment information and treatment result information of the reference patient are displayed on display 35 (step S15). In detail, a patient database 401 is stored and prepared in storage unit 40, such as an external server. In patient database 401, the results of pharmacological efficacy evaluation tests, analysis results of multiple immunostaining, treatment information, and treatment result information are associated with multiple patients for whom pharmacological efficacy evaluation test data and multiple immunostaining data were previously obtained and who underwent treatment.

[0045] The calculation unit 23 calculates (or may calculate in advance) a multidimensional vector for each of the multiple patients, with variables including the IC50 value of each drug in a drug efficacy evaluation test and the proportion of each type of immune cell in multiple immunostaining. The multidimensional vector for the target patient is also calculated. Next, the Euclidean distance between the position indicated by the multidimensional vector for each of the multiple patients and the position indicated by the multidimensional vector for the target patient is calculated, and the patient for whom the distance is less than a predetermined value is identified as the reference patient. Information on the actual treatments performed by the reference patient (including medications taken) and treatment result information are then displayed on the display 35. The treatment result information includes, for example, information indicating the effectiveness of the medications taken and the survival time of the reference patient. In the example of FIG. 6, the relationship between the survival rate and the number of days for each treatment method for the multiple reference patients is shown as treatment result information.

[0046] As described above, the disease evaluation support method includes the steps of: obtaining data on a pharmacological efficacy evaluation test for a predetermined drug using cells three-dimensionally cultured from a portion of the diseased tissue of a subject patient (step S11); obtaining data on multiplex immunostaining of a tissue specimen prepared from the diseased tissue of the subject patient (step S12); and displaying the results of the pharmacological efficacy evaluation test and the analysis results of the multiplex immunostaining side by side on the display 35 (step S13). This makes it possible to accurately grasp the state of the diseased tissue and predict the effects of a therapeutic drug. In other words, the disease evaluation support method can support a doctor in appropriately evaluating the disease of a subject patient. The same applies when, in step S11, data on a pharmacological efficacy evaluation test is obtained using cells three-dimensionally cultured from circulating cancer cells of the subject patient.

[0047] Drug efficacy evaluation tests are effective for selecting drugs that act on an unspecified number of cells (e.g., conventional anticancer drugs), but because the culture process itself takes time, there is a limit to the number of drugs that can be evaluated before treatment begins. Multiple immunostaining methods are particularly effective for selecting drugs that are effective specifically on cells in a specific state, but for other drugs, they only provide supplementary information on the disease state. In contrast, the disease evaluation support method described above, which displays the results of drug efficacy evaluation tests alongside the results of multiple immunostaining analysis, makes it possible to examine the effectiveness of various drugs and select appropriate therapeutic drugs and treatments tailored to the target patient.

[0048] Preferably, the region of the diseased tissue used for three-dimensional culture and the region of the diseased tissue used to prepare the tissue specimen for multiple immunostaining are close to each other, which allows the drug efficacy evaluation test data 211 and the multiple immunostaining data 212 to be obtained for substantially the same region, thereby enabling the selection of a more appropriate therapeutic drug or treatment.

[0049] Preferably, the disease evaluation support method further comprises a step (step S14) of determining an evaluation value for a predetermined drug using the results of the drug efficacy evaluation test and the analysis results of multiple immunostaining, and displaying the evaluation value on the display 35. In this way, displaying an evaluation value based on both the results of the drug efficacy evaluation test and the analysis results of multiple immunostaining enables the selection of a more appropriate therapeutic drug.

[0050] Preferably, the results of the pharmacological efficacy evaluation test, the analysis results of multiple immunostaining, treatment information, and treatment outcome information for a plurality of patients are prepared in advance. The disease evaluation support method further includes a step (step S15) of identifying a patient from the plurality of patients whose results of the pharmacological efficacy evaluation test and the analysis results of multiple immunostaining are similar to those of the target patient as a reference patient, and displaying the treatment information and treatment outcome information of the reference patient on the display 35. In this way, by displaying information on other patients whose disease states, etc. are similar to those of the target patient, it becomes possible to select a more appropriate therapeutic drug or treatment.

[0051] As described above, in step S11, it is preferable that efficacy evaluation tests for a plurality of types of drugs are performed simultaneously using the imaging device 41. This allows efficient performance of efficacy evaluation tests for a plurality of drugs. It is also preferable that efficacy evaluation tests for a plurality of concentrations of a predetermined drug are performed simultaneously using the imaging device 41. This allows efficient performance of efficacy evaluation tests for a plurality of concentrations of a drug.

[0052] Preferably, the analysis results of the multiple immunostaining include the analysis results of immune cells and tumor cells. This allows the relationship between immune cells and tumor cells to be confirmed, and the state of the diseased tissue (tumor tissue) to be appropriately understood. Furthermore, since the analysis results of the multiple immunostaining include the proportion or number of each type of immune cell, the state of the diseased tissue can be easily understood. Furthermore, the analysis results of the multiple immunostaining include the analysis results of immune cells in the tumor and various surrounding regions. This allows the positional relationship of immune cells to the tumor, i.e., spatial information about immune cells, to be easily understood. If the analysis results of the multiple immunostaining include a prediction of the effect of at least one type of drug, more appropriate therapeutic agents and treatments can be selected.

[0053] The memory unit 21 of the disease evaluation support device 1 described above stores data on a pharmacological efficacy evaluation test for a specific drug using cells three-dimensionally cultured from a portion of the diseased tissue of the target patient, and data on multiplex immunostaining of a tissue specimen prepared from the diseased tissue of the target patient. The display control unit 22 then displays the results of the pharmacological efficacy evaluation test and the analysis results of the multiplex immunostaining side by side on the display 35. This supports appropriate evaluation of the disease of the target patient and enables selection of an appropriate therapeutic drug or treatment tailored to the target patient. The same applies when obtaining data on a pharmacological efficacy evaluation test using cells three-dimensionally cultured from the target patient's circulating cancer cells.

[0054] The disease evaluation support method and disease evaluation support device 1 described above can be modified in various ways.

[0055] Depending on the type of disease of the subject patient, the analysis results of multiple immunostaining may include the analysis results of only either immune cells or tumor cells. The analysis results of multiple immunostaining preferably include the analysis results of immune cells and / or tumor cells.

[0056] Multiple immunostaining may be performed without using the immunostaining apparatus 42. In this case, the staining work is performed by an operator, and the sample is digitized using a specimen scanner (which may be used in combination with the imaging apparatus 41). The digital data (multiple immunostaining data 212) is then subjected to image analysis in the computer 3. Alternatively, the analysis results of the multiple immunostaining may be obtained by an external device (for example, a calculation unit of the immunostaining apparatus 42), and the analysis results may be stored in the storage unit 21 as multiple immunostaining data 212.

[0057] In the process of FIG. 3 , the step of obtaining data from a pharmacological efficacy evaluation test (step S11) and the step of obtaining data from multiple immunostaining (step S12) do not necessarily need to be performed in parallel (substantially simultaneously); the two steps may be performed at intervals. For example, a biopsy is performed in the early stages of cancer, and only multiple immunostaining is performed (step S12), followed by treatment. Later, as the cancer progresses, another biopsy is performed, and a pharmacological efficacy evaluation test using cultured cells is performed (step S11). The analysis results of the initial multiple immunostaining and the latest results of the pharmacological efficacy evaluation test using cultured cells are displayed side by side on the display 35 (step S13). This also supports appropriate evaluation of the patient's disease. The pharmacological efficacy evaluation test may be performed before the multiple immunostaining. Furthermore, when the amount of tissue to be collected is small, the pharmacological efficacy evaluation test and the multiple immunostaining may be performed at intervals. The diseased tissue sites used for the pharmacological efficacy evaluation test and the multiple immunostaining may be separated in space and / or time.

[0058] In the disease evaluation support method (and disease evaluation support device 1) described above, the display of the evaluation value of each drug in step S14 may be omitted. The same applies to the display of the treatment information and treatment result information of the reference patient in step S15. Furthermore, the display of the results of the pharmacological efficacy evaluation test and the analysis results of multiple immunostaining in step S13 may be omitted, and step S14 and / or step S15 may be performed following steps S11 and S12.

[0059] The disease evaluation support method and disease evaluation support device 1 can be used when the target patient has an infection, inflammation, or the like, in addition to cancer.

[0060] The configurations in the above-described embodiment and each modification may be combined as appropriate as long as they are not mutually contradictory. [Explanation of symbols]

[0061] 1. Disease evaluation support device 21 Memory section 22 Display control unit 35 Display 41 Imaging equipment 211 Drug Efficacy Evaluation Test Data 212 Multiplex immunostaining data 401 Patient Database Steps S11 to S15

Claims

1. A disease evaluation support method, comprising: a) obtaining data on efficacy evaluation tests of a predetermined drug using cells obtained by three-dimensionally culturing a portion of the diseased tissue of a subject patient; b) obtaining data on multiplex immunostaining of a tissue specimen prepared from the diseased tissue of the subject patient; c) displaying the results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display; Equipped with A disease evaluation support method, wherein the results of the drug efficacy evaluation test displayed on the display include the relationship between the concentration of the specified drug and the cell survival rate.

2. The disease evaluation support method according to claim 1, A disease evaluation support method, characterized in that the diseased tissue is cancer tissue.

3. 3. The disease evaluation support method according to claim 1 or 2, A disease evaluation support method, wherein the portion of the diseased tissue in step a) and the site of the diseased tissue used to prepare the tissue specimen in step b) are adjacent to each other.

4. A disease evaluation support method, comprising: a) obtaining data on efficacy evaluation of a predetermined drug using three-dimensionally cultured blood circulating cancer cells of a subject patient; b) obtaining data on multiplex immunostaining of a tissue specimen prepared from cancer tissue, which is a diseased tissue of the subject patient; c) displaying the results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display; Equipped with A disease evaluation support method, wherein the results of the drug efficacy evaluation test displayed on the display include the relationship between the concentration of the specified drug and the cell survival rate.

5. 5. A disease evaluation support method according to claim 1, comprising: The disease evaluation support method further comprises a step of determining an evaluation value of the specified drug using the results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining, and displaying the evaluation value on the display.

6. 6. A disease evaluation support method according to claim 1, comprising: the results of the drug efficacy evaluation test, the analysis results of the multiple immunostaining, treatment information, and treatment result information for a plurality of patients are prepared in advance; The disease evaluation support method comprises: a step of identifying, from among the plurality of patients, a patient whose results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining are similar to those of the target patient as a reference patient, and displaying the treatment information and the treatment result information of the reference patient on the display.

7. 7. A disease evaluation support method according to claim 1, comprising: A disease evaluation support method, characterized in that in the step a), the efficacy evaluation tests for a plurality of types of drugs are simultaneously performed using an imaging device.

8. 8. A disease evaluation support method according to claim 1, comprising: A disease evaluation support method, characterized in that in the step a), the efficacy evaluation tests for a plurality of concentrations of the predetermined drug are simultaneously performed using an imaging device.

9. 9. A disease evaluation support method according to claim 1, comprising: A disease evaluation support method, wherein the analysis results of the multiple immunostaining include analysis results of immune cells and / or tumor cells.

10. The disease evaluation support method according to claim 9, A disease evaluation support method, characterized in that the analysis results of the multiplex immunostaining include the proportion or number of each type of immune cell.

11. The disease evaluation support method according to claim 9 or 10, A disease evaluation support method, characterized in that the analysis results of the multiplex immunostaining include analysis results of immune cells in the tumor and various surrounding areas.

12. 12. A disease evaluation support method according to claim 1, comprising: A disease evaluation support method, characterized in that the analysis results of the multiple immunostaining include a prediction of the effect of at least one type of drug.

13. A disease evaluation support device, comprising: a memory unit that stores data on a drug efficacy evaluation test for a predetermined drug using cells obtained by three-dimensionally culturing a portion of a diseased tissue of a subject patient, and data on multiplex immunostaining of a tissue specimen prepared from the diseased tissue of the subject patient; a display control unit that displays the results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display; Equipped with A disease evaluation support device, wherein the results of the drug efficacy evaluation test displayed on the display include a relationship between the concentration of the predetermined drug and cell survival rate.

14. A disease evaluation support device, comprising: a memory unit that stores data on efficacy evaluation tests of a predetermined drug using three-dimensionally cultured blood circulating cancer cells of a subject patient, and data on multiplex immunostaining of a tissue specimen prepared from the cancer tissue, which is a diseased tissue of the subject patient; a display control unit that displays the results of the drug efficacy evaluation test and the analysis results of the multiple immunostaining side by side on a display; Equipped with A disease evaluation support device, wherein the results of the drug efficacy evaluation test displayed on the display include a relationship between the concentration of the predetermined drug and cell survival rate.

Images